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Indicators of a
Clandestine Laboratory

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GoalOfEarlyDetection
Earlydetectionofabiologicalagentallows:

Treatmentplanstobeimplementedwithin
therapeuticwindows
ProvidesinformationforPublicHealth
planningand;
Guidescommunityresponse

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Indicators of a Covert Biological
Attack
Suspicious material found at an unusual
location or time
Unusual sick, dead, or dying animals
Unusual illness for region or area
Definite pattern inconsistent with natural
disease

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 Indicators of a
Biological Attack
Unusual liquid, spray, or vapour, or
suspicious spraying devices or packages
Unusual swarms of insects

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 Biological vs Chemical Attack
Chemical incidents-
Characterized by the rapid onset of medical
symptoms (minutes to hours)
Easily observed signatures (colored residue, dead
foliage, pungent odor, and dead insect and animal life)
Biological incidents-
The onset of symptoms requires days to weeks and
there typically will be no characteristic signatures
Because of the delayed onset of symptoms in a
biological incident, the area affected may be greater
due to the migration of infected individuals
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 7

Covert Delivery
Aerosolise: Biological agents can be disseminated
using different aerosol methods, e.g. wet aerosol
methods- using garden sprayer or dry aerosol methods
where dry powder is dispersed eg, Anthrax attacks of
2001 in US -spores in letters

Put into food or water: Contamination of a city's water

supplies requires an unrealistically large amount of an
agent as well as introduction into the water after it
passes through a regional treatment facility.
 8

Injected into the skin,

either by insect vectors
such as fleas or ticks

An umbrella might be
ideal for assassination, as
in Markovs assassination
in London in 1978, but this
is not likely to be used to
cause mass casualties.
 9

Overt Delivery
Used in explosives (artillery, missiles, detonated bombs):
The attack is announced
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Covert vs Overt Event

Overt Covert
Recognition Early Delayed
Response Early Delayed
Treatment Early Delayed
Responders Usual First Health Care
Responders Workers
 Clandestine Laboratories

Clan labs
A hidden lab used to illegally grow or
produce biological agents.
E.g., bacteria or viruses.

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ClandestineLabIndicators

VS.

CommercialCentrifuge HomemadeCentrifuge

VS.

CommercialGlovebox HomemadeGlovebox
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VS.

Commercialautoclave Homemadeautoclave

VS.

Commercialincubator Homemadeincubator
IndicatorsofBacterialClandestineLab

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15
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Commerciallyavailablemediaandhomemademediausedto
growBACTERIA
IndicatorsofAViralClandestineLab

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 Growing Viruses
Cellsinflasksandwells
usedtogrowVIRUSES

CellsintidalflowbioreactorsforcommercialgrowthofVIRUSES
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Viral Lab Egg Cultures

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 Detection of
Biological Agents
PRESUMPTIVE Detection&Testingof
BiologicalAgents:
ProteinScreening
ProteinBasedPowderScreeningKits
Spectrometry
Antigen/antibodytests
HandHeldAssays(HHAs)/Biotickets
MolecularDetection
PolymeraseChainReaction(PCR)
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Protein-Based Powder Screening

Kits
Identifies the presence or
absence of protein in
powders
The results are read as
simple colour changes when
swab is placed in protein 20/20BioCheck
detection tube powderscreeningkit
 InfraredSpectroscopy

Primarilyachemicaldetection
technology
ComparestheInfraredspectrumof
anunknownpowderorliquid
againstadatabaselibraryofspectra
ofknownchemicals

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 Protein Detection - Advantages

Technically simple to perform in field

Results in as little as 5 minutes

The results are simple colour changes in the

case of powder kits or a protein warning in the
case of spectroscopy

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Protein Detection - Disadvantages

Provides info on protein content ONLY!

Poor specificity
Poor sensitivity
What does a positive result mean??
What does a negative result mean?
Antigen antibody tests HHAs / Biotickets

Thetestkitisloadedwithknown antibodies
Whenasampleantigenisaddedtothesample
port,theantigenbindswiththeantibodiescreating
acolour change

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Antigen antibody tests HHAs / Biotickets

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 The Interaction between an Antigen and Antibody :-

Antigen must FIT the Antibody

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Howanantigen/antibodytestworks:

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CommonAntigen/Antibodytest:

Mostcommonandsuccessfulbioticket isthe
pregnancytest

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Antigen/AntibodyTests
Advantages:
Canbeveryfast
Easytouse
Portable
Canbeveryspecificand
sensitive

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Antigen/AntibodyTests
Disadvantages:
Inhibitedbycontaminants
Falsenegativeresultsmayoccur:
Antigenbelowthedetectableconcentration
Overloadingofthetest
Falsepositiveresultsmayoccur:
Unknowncrossreactions
Testsaretemperaturedependant

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MolecularDetection
PolymeraseChainReaction(PCR)

Called molecular photocopying

Technique used to amplify small segments
of DNA in order to identify the organism or
agent
Mimics the normal process that takes
place in your cells when they divide

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How does PCR work?
Polymerase Chain Reaction (PCR)
Mimics DNA Replication: a normal process where
cells copy DNA to divide
Copies specific fragments of DNA to detectable levels

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Real Time PCR: Results

Positive
threshold

10-fold dilution series of target DNA

Iffluorescentsignalcrossesthreshold,sampleispositive
Quantitativedetectionobserved
106 copiesoftargetDNAdetectedatcycle20
105 copiesoftargetDNAdetectedatcycle23
 36

MolecularDetection:
AdvantagesPCRbasedassays
1. Sensitive
AslittleasONEDNACOPYdetected

2. Specific
Primerstargetuniquesequenceofinterest
Probes(realtime)enhancespecificity

3. Quick(realtime)
~3060min)

4. Quantitative (realtime)
IndicatesstartingamountofDNAtemplateinsample

5. Practicalforthefield
Ex:SmartCycler,Lightcycler,RAZORRealTimePCRinstruments
MolecularDetection:
DisadvantagesofPCR

Qualified&trainedtechnicianneedtorun
andinterpretresults
Environmentalcontaminationofsample
caninhibitresults
Equipmentisexpensive

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RAZOR System: Real-Time PCR

DetectsandidentifiesBTagentsusingRealTimePCR
anthrax,ricin,smallpox,plague,avianinfluenza,botulism,plague,
tularemia
Createdforfirstrespondersandfrontlinemilitarytroops
Batterypowered;Portable
Pouchescontainallreagentsforsamplepurification&PCR
12samplesperrun
Resultsin~45minutes
 39

RAZOR System Procedure

RAZOR
PCR machine

Pouch

1. TurnonRAZOR,selecttest
2.Loadsyringewithtestsample
3. Insertthesyringeintotheinletport
4. Allowthesampletobeautomaticallypulledintothepouch
5.InsertpouchintoslotofRAZOR,startrun
Why is it important to send all samples
to the appropriate labs for definitive
testing?

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 The Canadian Bioterrorism Laboratory
Response Network
LaboratoryResponse
NetworkLabs(westto ReferenceLab
east): NationalReference
VancouverBC
EdmontonAB
ReginaSK
TorontoONT
SteAnnedeBellevueQUE
St.John,NB(2014?)

NationalReference
Laboratory:
NML(Wpg,CL3&4,civilian)
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Questions?

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